Objective To explore the influences of extracellular matrices (ECM) secreted by ultraviolet B (UVB)-induced senescent fibroblasts on the proliferation of and extracellular signal-regulated kinase (ERK) signaling in HaCaT cells. Methods Fibroblasts were irradiated with UVB of 15 mJ/cm2 once daily for 5 days to induce premature senescence, which was identified by SA-β-gal staining 72 hours after the last irradiation.HaCaT cells were divided into 3 groups and inoculated into plates coated with extracellular matrices secreted by non-senescent (PRE-ECM) or senescent fibroblasts (SIPS-ECM) or into uncoated plates (NON-ECM), fol-lowed by additional culture. U0126, an inhibitor of ERK1/2, was used to treat the HaCaT cells 1 hour before inoculation. Then, MTT assay was carried out to detect the proliferation of HaCaT cells after a 3-day culture,Western blot to assess the phosphorylation of ERK at 0.5, 1, 2 and 4 hours after the inoculation, flow cytometry to analyse cell cycle and apoptosis after 24 hours of culture. Results The most rapid and intense phosphory-lation of ERK was observed in SIPS-ECM group. Inhibiting the activation of ERK pathway with U0126 could completely suppress the promoting effect of ECM from senescent fibroblasts on the proliferation of HaCaT cells.After the blocking of ERK activation, the proportion of HaCaT cells in S and G2/M phase decreased from 37.40%, 41.34% and 43.31% to 29.41%, 36.48% and 39.96%, respectively, in NON-ECM, PRE-ECM and SCIP-ECM group. Conclusion The ECM produced by UVB-induced senescent fibroblasts promote the prolifera-tion of HaCaT cells via inducing the phosphorylation of ERK.

Background and purpose:Risk of invasive cervical carcinoma (ICC) and its precancerous lesions following high risk human papillomavirus (hrHPV) infection may vary according to HPV types and geographic regions. Analyzing HPV-type distribution in cervical samples from local women aged 30 years and older, this study aimed to identify HPV types with higher risk of developing CIN2+, and to compare diagnostic performance for CIN2+using these types and type combinations. Methods:Cervical samples with histology follow-up from patients of a tertiary cancer center in Shanghai were collected for HPV genotyping by PCR-RDB. The risk associations of HPV types with CIN2+were estimated by logistic regression analysis, and ROC curves were plotted for diagnostic performance evaluation. Results:A total number of 413 specimens were obtained, including 38 CIN1, 184 CIN2/3, 126 ICC patients and 65 negative control people. The 4 most common HPV types in CIN2+were HPV16, 58, 33 and 18, in descending order. And only HPV16 (P<0.000 1), 58 (P=0.002), 33 (P=0.015) were signiifcantly associated with CIN2+lesions. Besides, the area under the ROC curve of the HPV16/18/33/58 test scored statistically higher than the HPV16/18 test did (P=0.006 6). Conclusion:A combined test of HPV16/18/33/58 may offer better performance for detecting CIN2+lesions in our geographic region.

OBJECTIVETo investigate the clinical results of pedicle-screw placement and reduction on the level of injured vertebrae in treating thoracolumbar fractures.

METHODSFrom March 2006 to May 2009, 27 patients with thoracolumbar fractures were treated with short-segment fixation system and pedicle-screw placement on injured vertebrae through posterior approach. There were 19 males and 8 females with an average age of 36 years old ranging from 24 to 68 years. Eleven patients were injuried by traffic accident, 9 patients were injuried by falling from high, 5 patients were injuried by crush from weighty object, 2 patients were others injuries. Thoracolumbar fractures were classified according to Gertibein classification: 2 cases were of type A1, 3 cases A2, 12 cases A3, 5 cases B1, 5 cases B2. Among them, 14 cases were accompanied with spinal cord injuries,functional assessment of nerves was assessed according to Frankel criteria: 5 cases were of grade B, 3 cases C, 6 cases D. Lumbago according to Denis standard, grade P5 was in all cases. The injured level of 5 cases was in T11, 11 cases in T12, 8 cases in L1, 3 cases in L2. From injury to operation it was 2-15 days with an average of 4 days. The ratio of vertebral height between anterior and posterior border, anteroposterior Cobb angle were compared before and after operation. Function of nerves and index of lumbago were evaluated by Frankel and Denis classification.

RESULTSAll patients were followed up from 14 to 17 months with the mean of 15.5 months. At final follow-up,according to Frankel score system to evaluate function of nerve,2 cases were of grade B, 4 case grade C, 3 cases grade D, 5 cases grade E; according to Denis score system to evaluate index of lumbago, there was P1 (without pain) in 17 cases, P2 (between whiles minute pain, need not treatment) in 8 cases, P3 (moderate pain, between whiles need medication) in 2 cases. The ratio of vertebral height between anterior and posterior border from preoperatively 0.32 +/- 0.14 to postoperatively 0.85 +/- 0.03 (P<0.01); anteroposterior Cobb angle from preoperatively (25.3 +/- 7.8) degrees to postoperatively (11.1 +/- 1.5) degrees (P<0.01). At final follow-up, the ratio of vertebral height between anterior and posterior border, anteroposterior Cobb angle were respectively 0.81 +/- 0.06 and (11.8 +/- 1.9) degrees, there was no significant difference between postoperative and at final follow-up (P>0.05). No complications such as infection, internal fixation failure,nerve injuries were found during follow-up.

CONCLUSIONPosterior short-segment fixation system and pedicle-screw placement on level of injured vertebrae is one kind of effective method in treating thoracolumbar fracture, which can maintain corrective effect and obtain better clinical outcomes.

Adult ; Aged ; Bone Screws ; Female ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Spinal Fractures ; surgery ; Thoracic Injuries ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the effect of hydrogen dioxide (H(2)O(2)) on the expression of vascular endothelial growth factor (VEGF) in human bronchiolar epithelial (HBE) cells.

METHODSMTT assay was used to assess HBE cell viability after exposure to different concentrations of H(2)O(2). VEGF/beta-actin gene fragments were amplified simultaneously by RT-PCR from the total HBE cell RNA, and VEGF protein expression in the cells was detected using ELISA.

RESULTSThe exposure to 200 micromol/L H(2)O(2) did not obviously affected the cell viability. Compared with those in the control cell, VEGF165/beta-actin and VEGF189/beta-actin ratios were significantly increased in the cells after treatment with 50, 200, and 600 micromol/L H(2)O(2) (P<0.05). The protein expression of VEGF significantly increased after 50 micromol/L H(2)O(2) treatment (P<0.05), but significantly decreased with pretreatment with the PI3K inhibitor Ly294002 (P>0.05).

CONCLUSIONOxidative stress increases the expression of VEGF via a PI3K-dependent pathway in human bronchiolar epithelial cells, which may play an important role in the onset and maintenance of chronic inflammation in asthma.

Actins ; metabolism ; Bronchi ; cytology ; Cell Line ; Epithelial Cells ; cytology ; metabolism ; Humans ; Hydrogen Peroxide ; pharmacology ; Oxidative Stress ; physiology ; Phosphatidylinositol 3-Kinases ; metabolism ; Signal Transduction ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

BACKGROUNDRecent studies have shown that T helper type-2 (Th2) cells can induce the apoptosis of CD4+CD25+ Treg cells or resist the immunosuppressive effect of Treg cells. We hypothesize that an imbalance of Th2/Treg is present in patients with allergic asthma.

METHODSTwenty-two patients with mild asthma, 17 patients with moderate to severe asthma, and 20 healthy donors were enrolled. All patients were allergic to house dust mites. The proportion of peripheral blood CD4+CD25+ Treg cells and Th2 cells were determined by flow cytometry. The concentration of interleukin (IL)-10, transforming growth factor (TGF)-β and IL-4 in plasma was determined by enzyme linked immunosorbent assay. In these subjects, peripheral blood mononuclear cells from 17 mild asthmatic patients, 13 moderate to severe asthmatic patients and 14 healthy donors were acquired and expression of forkhead box P3 (Foxp3) and GATA-3 mRNA was detected by reverse-transcriptase polymerase chain reaction.

RESULTSCompared with healthy donors and patients with mild asthma, the percent of CD4+CD25+ Treg cells and plasma IL-10 levels were decreased in patients with moderate to severe asthma. There were no significant differences in Foxp3 mRNA expression among three groups, but a downward trend seen among patients with asthma. However, the percent of Th2 cells, IL-4 levels and expression of GATA-3 mRNA was markedly higher in patients with mild and moderate to severe asthma than in the control group. The ratio of Th2/Treg and their cytokines was increased in allergic asthma, especially for moderate to severe asthma. The ratio of GATA-3/Foxp3 mRNA was also increased in allergic asthma. In patients with moderate to severe asthma, the percentage of peripheral blood Treg cells was negatively correlated to the percentage of Th2 cells and IL-4 levels.

CONCLUSIONSThe decline of CD4+CD25+ Treg cells in patients with moderate to severe asthma may play an important role in progress of the disease. Furthermore, the deficiency of CD4+CD25+ Treg cells was associated with the over-expression of Th2 response.

Asthma ; etiology ; immunology ; Cytokines ; blood ; Forkhead Transcription Factors ; genetics ; GATA3 Transcription Factor ; genetics ; Humans ; RNA, Messenger ; analysis ; T-Lymphocytes, Regulatory ; immunology ; Th2 Cells ; immunology

OBJECTIVETo examine the association between polymorphism of vascular endothelial growth factor(VEGF)1498 C/T,936 C/T and colorectal adenoma genetic susceptibility.

METHODSA case-control study of 224 colorectal adenomas and 200 controls was conducted and VEGF genotypes were determined based on TaqMan-probe assay. The epidemiological factors were collected through questionnaire. Accordingly, the clinicopathological data of each sample were also investigated.

RESULTSThe carriage of 936 CT and CT+TT genotypes had significantly higher risk of colorectal adenoma (CT vs. CC, OR=2.00, 95% CI: 1.23-3.25, P=0.006; CT+TT vs. CC, OR=2.04, 95% CI:1.28-3.26, P=0.003). 936-T allele carriage had increased risk of colorectal adenoma (OR=1.91, 95% CI:1.25-2.91, P=0.003). The genotypes of 1498 C/T and the frequency of C/T allele showed no differences between healthy persons and patients (P>0.05). In patients with 936 CT+TT and 936-T allele implied a tendency of villous adenoma category (CT+TT vs. CC, OR=2.54, 95% CI:1.12-5.75, P=0.040; T allele vs. C allele, OR=3.08, 95% CI, 1.64-5.80, P=0.001).

CONCLUSIONVEGF 936 C/T polymorphism can influence susceptibility to colorectal adenoma.

Adenoma ; genetics ; Adult ; Case-Control Studies ; Colorectal Neoplasms ; genetics ; Female ; Genetic Predisposition to Disease ; Genotype ; Humans ; Middle Aged ; Polymorphism, Single Nucleotide ; Vascular Endothelial Growth Factor A ; genetics

Cell Line, Tumor ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Humans ; Microbubbles ; Oligoribonucleotides, Antisense ; genetics ; Transfection ; methods ; Ultrasonics

OBJECTIVETo investigate the properties of screw-bone interface of expansive pedicle screw (EPS) in osteoporotic sheep by micro-CT and histological observation.

METHODSSix female sheep with bilateral ovariectomy-induced osteoporosis were employed in this experiment. After EPS insertion in each femoral condyle, the sheep were randomly divided into two groups: 3 sheep were bred for 3 months (Group A), while the other 3 were bred for 6 months (Group B). After the animals being killed, the femoral condyles with EPS were obtained, which were three-dimensionally-imaged and reconstructed by micro-CT. Histological evaluation was made thereafter.

RESULTSThe trabecular microstructure was denser at the screw-bone interface than in the distant parts in expansive section, especially within the spiral marking. In the non-expansive section, however, there was no significant difference between the interface and the distant parts. The regions of interest (ROI) adjacent to EPS were reconstructed and analyzed by micro-CT with the same thresholds. The three-dimensional (3-D) parameters, including tissue mineral density (TMD), bone volume fraction (BVF, BV/TV), bone surface/bone volume (BS/BV) ratio, trabecular thickness (Tb.Th), and trabecular separation (Tb.Sp), were significantly better in expansive sections than non-expansive sections (P less than 0.05). Histologically, newly-formed bony trabeculae crawled along the expansive fissures and into the center of EPS. The newly-formed bones, as well as the bones at the bone-screw interface, closely contacted with the EPS and constructed four compartments.

CONCLUSIONSThe findings of the current study, based on micro-CT and histological evaluation, suggest that EPS can significantly provide stabilization in osteoporotic cancellous bones.

Animals ; Bone Screws ; Bone and Bones ; diagnostic imaging ; pathology ; Female ; Osteoporosis ; diagnostic imaging ; pathology ; Sheep ; Tomography, X-Ray Computed

Adolescent ; Anemia, Hemolytic ; etiology ; Female ; Hemolysis ; Hepatic Encephalopathy ; etiology ; Hepatolenticular Degeneration ; complications ; Humans

OBJECTIVETo investigate the frequency and clinical implication of JAK2 mutation in patients with myeloproliferative neoplasm(MPN)and the correlation between the mutation and thrombosis.

METHODSThe clinical and laboratory data of 107 MPN patients was retrospectively analyzed. JAK2 mutation were detected with allele-specific polymerase chain reaction (AS-PCR) and sequencing. The significance of the mutation in disease diagnosis and molecular pathogenesis and the correlation between the mutation and thrombosis was analysed.

RESULTSJAK2 mutation was detected in 71 (66.4%) and thrombosis in 34 (31.8%) of the 107 MPN patients. Thrombosis occurred in 34.8% (16/46) of polycythemia vera (PV), 32.6% (14/43) of essential thrombocythemia (ET), and 22.2% (4/18) of primany myelofibrosis (PMF) patients. The difference among the 3 groups was not significant (χ(2) = 0.96, P > 0.05). The frequency of thrombosis in JAK2(+) MPN patients (82.4%, 28/34) was higher than that in JAK2(-) MPN patients (17.6%, 6/34) (χ(2) = 5.71, P < 0.05). The frequency of thrombosis in MPN patients > 60 years was higher (41.5%, 27/65) than that in patients < 60 years (16.7%, 7/42) (χ(2) = 7.28, P < 0.01).

CONCLUSIONJAK2 V617F mutation occurs in significant percentage of Chinese patients with MPN. Patients with JAK2 mutation and older age are more succeptible to thrombosis. JAK2 mutation screening in patients with unknown thrombosis is helpful to reveal the underlying latent-MPN.

Humans ; Mutation ; Myeloproliferative Disorders ; genetics ; Neoplasms ; Thrombocythemia, Essential ; genetics ; Thrombosis